Xie Lihan, Li Weinan, Fan Shiqi, Jin Jinsong
College of Traditional Chinese Medicine, Hubei University of Chinese Medicine, Wuhan, Hubei, China.
Department of Nephrology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
J Biochem Mol Toxicol. 2025 Sep;39(9):e70455. doi: 10.1002/jbt.70455.
Diabetic kidney disease (DKD) is a common complication of diabetes, often characterized by podocyte injury, proteinuria, and eventual renal failure. Sinomenine hydrochloride (SH), an active component derived from traditional Chinese medicine, is clinically effective in treating kidney diseases. This study investigates the protective effects of SH on podocytes under high-glucose conditions and its mechanism of action. Mouse podocytes (MPC-5) were treated with SH at concentrations of 50, 200, and 600 μg/mL under high-glucose conditions (30 mmol/L) for 24 h to establish a DKD model. Cell viability was assessed using CCK-8 assays, and apoptosis rates were measured using flow cytometry. Autophagy levels were evaluated by detecting LC3-II, Beclin-1, and P62 proteins via Western blot analysis, while the involvement of the PI3K/AKT/mTOR pathway was analyzed by examining phosphorylated AKT and mTOR. Transmission electron microscopy was employed to observe autophagosomes. SH improved podocyte viability, reduced apoptosis, and enhanced autophagic activity by increasing LC3-II and Beclin-1 expression while decreasing P62 levels. SH also downregulated p-AKT and p-mTOR, indicating inhibition of the PI3K/AKT/mTOR pathway. Electron microscopy confirmed increased autophagosomes in SH-treated groups. SH protects podocytes in a high-glucose environment by enhancing autophagy through inhibition of the PI3K/AKT/mTOR pathway. These findings provide insights into SH as a potential therapeutic agent for DKD management.
糖尿病肾病（DKD）是糖尿病常见的并发症，其特征通常为足细胞损伤、蛋白尿及最终的肾衰竭。盐酸青藤碱（SH）是一种源自中药的活性成分，在治疗肾脏疾病方面具有临床疗效。本研究探讨了SH在高糖条件下对足细胞的保护作用及其作用机制。在高糖条件（30 mmol/L）下，用浓度为50、200和600 μg/mL的SH处理小鼠足细胞（MPC-5）24小时，以建立DKD模型。使用CCK-8法评估细胞活力，采用流式细胞术检测凋亡率。通过蛋白质免疫印迹分析检测LC3-II、Beclin-1和P62蛋白来评估自噬水平，同时通过检测磷酸化的AKT和mTOR来分析PI3K/AKT/mTOR通路的参与情况。采用透射电子显微镜观察自噬体。SH通过增加LC3-II和Beclin-1表达同时降低P62水平，改善了足细胞活力，减少了凋亡，并增强了自噬活性。SH还下调了p-AKT和p-mTOR，表明其抑制了PI3K/AKT/mTOR通路。电子显微镜证实SH处理组的自噬体增加。SH通过抑制PI3K/AKT/mTOR通路增强自噬，从而在高糖环境中保护足细胞。这些发现为SH作为DKD治疗的潜在治疗药物提供了见解。
